Transcriptome Analyses of In Vitro Exercise Models by Clenbuterol Supplementation or Electrical Pulse Stimulation

Exercise has beneficial effects on human health and is affected by two different pathways; motoneuron and endocrine. For the advancement of exercise research, in vitro exercise models are essential. We established two in vitro exercise models using C2C12 myotubes; EPS (electrical pulse stimulation) for a motoneuron model and clenbuterol, a specific β2 adrenergic receptor agonist, treatment for an endocrine model. For clenbuterol treatment, we found that Ppargc1a was induced only in low glucose media (1 mg/mL) using a 1-h treatment of 30 ng/mL clenbuterol. Global transcriptional changes of clenbuterol treatment were analyzed by RNA-seq and gene ontology analyses and indicated that mitogenesis and the PI3K-Akt pathway were enhanced, which is consistent with the effects of exercise. Cxcl1 and Cxcl5 were identified as candidate myokines induced by adrenaline. As for the EPS model, we compared 1 Hz of 1-pulse EPS and 1 Hz of 10-pulse EPS for 24 h and determined Myh gene expressions. Ten-pulse EPS induced higher Myh2 and Myh7 expression. Global transcriptional changes of 10-pulse EPS were also analyzed using RNA-seq, and gene ontology analyses indicated that CaMK signaling and hypertrophy pathways were enhanced, which is also consistent with the effects of exercise. In this paper, we provided two transcriptome results of in vitro exercise models and these databases will contribute to advances in exercise research

Star Formation Timescales of the Halo Populations from Asteroseismology and Chemical Abundances

We combine asteroseismology, optical high-resolution spectroscopy, and kinematic analysis for 26 halo red giant branch stars in the \textit{Kepler} field in the range of $-2.5<[\mathrm{{Fe}/{H}}]<-0.6$. After applying theoretically motivated corrections to the seismic scaling relations, we obtain an average mass of $0.97\pm 0.03\,\mathrm{M_{\odot}}$ for our sample of halo stars. Although this maps into an age of $\sim 7\,\mathrm{Gyr}$, significantly younger than independent age estimates of the Milky Way stellar halo, we considerer this apparently young age is due to the overestimation of stellar mass in the scaling relations. There is no significant mass dispersion among lower red giant branch stars ($\log g>2$), which constrains a relative age dispersion to $<18\%$, corresponding to $<2\,\mathrm{Gyr}$. The precise chemical abundances allow us to separate the stars with [{Fe}/{H}]$>-1.7$ into two [{Mg}/{Fe}] groups. While [$\alpha$/{Fe}] and [{Eu}/{Mg}] ratios are different between the two subsamples, [$s$/Eu], where $s$ stands for Ba, La, Ce, and Nd, does not show a significant difference. These abundance ratios suggest that the chemical evolution of the low-Mg population is contributed by type~Ia supernovae, but not by low-to-intermediate mass asymptotic giant branch stars, providing a constraint on its star formation timescale as $100\,\mathrm{Myr}<\tau<300\,\mathrm{Myr}$. We also do not detect any significant mass difference between the two [{Mg}/{Fe}] groups, thus suggesting that their formation epochs are not separated by more than 1.5 Gyr.Comment: 44 pages. accepted versio

Protein kinase A-dependent substance P expression by pituitary adenylate cyclase-activating polypeptide in rat sensory neuronal cell line ND7/23 cells.

The neurotrophic effects of pituitary adenylate cyclase-activating polypeptide (PACAP) on rat sensory neuronal cell line ND7/23 cells were investigated. PACAP caused a concentration-dependent increase in the number of neurite-bearing cells and the expression of the substance P precursor (PPT) mRNA in 24 h. The effects of PACAP were mimicked by vasoactive intestinal polypeptide with lower potency and dibutyryl-cyclic AMP, and inhibited by inhibitors of protein kinase A, ERK kinase or p38 kinase, KT5720, U0126, or SB203580, respectively. In a PPT promoter luciferase reporter assay, the increase of PPT mRNA was the result of an increase in PPT gene transcriptional activity by PACAP. The increasing effects of PACAP on PPT mRNA were similarly observed in primary cultured rat dorsal root ganglion cells. Thus, PACAP could induce differentiation-like phenomena in sensory neurons in a cAMP-, protein kinase A-, ERK kinase-, and p38 kinase-dependent manner. These results provide evidence of the neurotrophic action of PACAP, which may function to rescue damaged neurons or to switch the neuronal phenotype in injured or inflamed sensory neurons.The neurotrophic effects of pituitary adenylate cyclase-activating polypeptide (PACAP) on rat sensory neuronal cell line ND7/23 cells were investigated. PACAP caused a concentration-dependent increase in the number of neurite-bearing cells and the expression of the substance P precursor (PPT) mRNA in 24 h. The effects of PACAP were mimicked by vasoactive intestinal polypeptide with lower potency and dibutyryl-cyclic AMP, and inhibited by inhibitors of protein kinase A, ERK kinase or p38 kinase, KT5720, U0126, or SB203580, respectively. In a PPT promoter luciferase reporter assay, the increase of PPT mRNA was the result of an increase in PPT gene transcriptional activity by PACAP. The increasing effects of PACAP on PPT mRNA were similarly observed in primary cultured rat dorsal root ganglion cells. Thus, PACAP could induce differentiation-like phenomena in sensory neurons in a cAMP-, protein kinase A-, ERK kinase-, and p38 kinase-dependent manner. These results provide evidence of the neurotrophic action of PACAP, which may function to rescue damaged neurons or to switch the neuronal phenotype in injured or inflamed sensory neurons

トウニョウビョウ ケア ノ リスク マネージメント

The number of diabetics has been increasing in recent years. The diabetics are under varioustreatments, including the improvement of life habit and the medication for diabetes with insulin.Our hospital set a team of diabetic care, which is composed of a diabetic specialist, certified diabeteseducators（CDEs）, nurses, dietricians and pharmacists. This team takes great care of the diabetics.For medical safety measures, the department of risk management was organized in our hospital.The department investigated the cases of Hiyari-Hatto within 1 year and 3 months, from 2005to 2006, and found that 3% of them was the diabetic case, which was caused by the nurses exceptCDEs. Therefore the department made the manual of diabetic therapy in cooperation with theCDEs. All the staffs in our hospital were educated by the seminars according to the manual. Theknowledge about the diabetic therapy proved to be mostly accurate one year after the last seminar.For the improvement of medical safety, the department of risk management helps the CDEswith holding the educational seminars by giving the informations after analyzing the cases of Hiyari-Hatto and the questionnaires following the seminars

Transcriptome Analyses of In Vitro Exercise Models by Clenbuterol Supplementation or Electrical Pulse Stimulation

Exercise has beneficial effects on human health and is affected by two different pathways; motoneuron and endocrine. For the advancement of exercise research, in vitro exercise models are essential. We established two in vitro exercise models using C2C12 myotubes; EPS (electrical pulse stimulation) for a motoneuron model and clenbuterol, a specific β2 adrenergic receptor agonist, treatment for an endocrine model. For clenbuterol treatment, we found that Ppargc1a was induced only in low glucose media (1 mg/mL) using a 1-h treatment of 30 ng/mL clenbuterol. Global transcriptional changes of clenbuterol treatment were analyzed by RNA-seq and gene ontology analyses and indicated that mitogenesis and the PI3K-Akt pathway were enhanced, which is consistent with the effects of exercise. Cxcl1 and Cxcl5 were identified as candidate myokines induced by adrenaline. As for the EPS model, we compared 1 Hz of 1-pulse EPS and 1 Hz of 10-pulse EPS for 24 h and determined Myh gene expressions. Ten-pulse EPS induced higher Myh2 and Myh7 expression. Global transcriptional changes of 10-pulse EPS were also analyzed using RNA-seq, and gene ontology analyses indicated that CaMK signaling and hypertrophy pathways were enhanced, which is also consistent with the effects of exercise. In this paper, we provided two transcriptome results of in vitro exercise models and these databases will contribute to advances in exercise research